The complete sequence (22 kilobases) of murine coronavirus gene 1 encoding the putative proteases and RNA polymerase

Lee, Han-Jung; Shieh, Chien-Kou; Gorbalenya, Alexander E.; Koonin, Eugene V.; Monica, Nicola La; Tuler, Jeremy; Bagdzhadzhyan, Anush; Lai, Michael M. C.

doi:10.1016/0042-6822(91)90071-i

Cited by 346 publications

(469 citation statements)

References 51 publications

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“…One of the initial steps in MHV replication is the translation of gene 1 from the input genomic RNA. Gene 1 encompasses approximately 21.8 kb at the 5' end of the genome and contains two overlapping large open reading frames (Lee et al, 1989;Bonilla et al, 1994). The ®rst open reading frame ORF1a encodes 4469 amino acids.…”

Section: Introductionmentioning

confidence: 99%

Localization of mouse hepatitis virus open reading frame 1A derived proteins

Bi¹,

Piñón²,

Hughes³

et al. 1998

J Neurovirol

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We have investigated the intracellular localization of proteolytic cleavage products encoded in the 5' portion of mouse hepatitis virus (MHV) gene 1. Immuno¯uorescent labeling of cells with an antiserum which recognizes p28, the ORF1a N-terminal cleavage product, resulted in widespread somewhat granular cytoplasmic staining, indicating that this protein is widely distributed in the cytoplasm of MHV-infected, but not control uninfected cells. Immunouorescent staining of infected cells with antisera which recognize the downstream polypeptides, p65, p240 and p290 labeled discrete vesicular perinuclear structures. Double immuno¯uorescent labeling of BHK cells expressing the MHV receptor (BHK MHVR1 ) and infected with MHV-A59 with a Golgi-speci®c anti-mannosidase II monoclonal antibody and with antiserum recognizing each of these anti-MHV ORF1a polypeptides, showed that the p240 and p290 polypeptides were localized in discrete vesicular structures that overlapped the Golgi complex. Labeling with antibodies speci®c for p65 colocalized with the Golgi region, and showed staining of the perinuclear cytoplasm as well. Plasmids containing sequences contained in the ®rst 6.75 kb of ORF1a have been expressed using the coupled vaccinia virus-T7 polymerase system. Immuno¯uorescent labeling of transfectants with the anti-ORF1a antisera showed patterns of antigen distribution similar to those observed in cells infected with MHV-A59. A deletion analysis with constructs containing only portions of the ORF1a sequence indicated that 303 amino acids containing the ®rst papain-like protease domain (PLP-1) was suf®cient to associate this protein with the Golgi.

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Section: Introductionmentioning

confidence: 99%

Localization of mouse hepatitis virus open reading frame 1A derived proteins

Bi¹,

Piñón²,

Hughes³

et al. 1998

J Neurovirol

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“…The murine coronavirus (mouse hepatitis virus, MHV) genome of 31 kb (Spaan et al, 1988;Lee et al, 1991) encodes four major structural proteins and several nonstructural proteins. The structural proteins are the nucleocapsid (N) protein of about 50K, the membrane (M) protein of 20K to 25K, the haemaggglutininesterase (HE) protein of about 65K and the spike (S) protein of 150K to 180K (Siddell et al, 1983;Spaan et al, 1988).…”

Section: Introductionmentioning

confidence: 99%

Molecular cloning and expression of a spike protein of neurovirulent murine coronavirus JHMV variant c1-2

Taguchi¹,

Ikeda²,

Shida³

1992

Journal of General Virology

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A cDNA encoding the spike (S) protein of the neurovirulent murine coronavirus JHMV variant cl-2 was isolated and sequenced. Analysis of the cDNA revealed that the S protein consists of 1376 amino acids, as does the S protein of mouse hepatitis virus 4. We inserted the cDNA into the genome of vaccinia virus to obtain a recombinant vaccinia virus (rVV). The S protein expressed in RK13 cells infected by the rVV was shown to be electrophoretically and immunologically indistinguishable from the S protein produced in DBT cells infected with cl-2 virus. RVV infection of rats and mice induced S protein-specific antibody production detectable by immunofluorescence and neutralization. Moreover, the S protein expressed by the rVV induced syncytium formation not only in mouse DBT and L cells, which are susceptible to cl-2 virus infection, but also in rabbit RK13 cells, which are not susceptible to cl-2 virus infection. This result suggests the possibility that RK13 cells have binding sites for the cl-2 virus S protein.

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“…A novel coronavirus, named SARS coronavirus (SARS-CoV), was soon identified as the etiological agent of the infection (1)(2)(3). It was revealed that the open reading frame of the replicase gene in SARS-CoV genome, similar to other coronaviruses (4,5), features the sequence motifs of both papainlike proteinase and 3C-like proteinase (3CL pro ). SARS 3CL pro is fully conserved among all released SARS coronavirus genome sequences and is highly homologous to other coronavirus 3CL proteinases (6,7).…”

mentioning

confidence: 99%

Severe Acute Respiratory Syndrome Coronavirus 3C-like Proteinase N Terminus Is Indispensable for Proteolytic Activity but Not for Enzyme Dimerization

Chen¹,

Chen²,

Tan³

et al. 2005

Journal of Biological Chemistry

123

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Severe acute respiratory syndrome (SARS) coronavirus is a novel human coronavirus and is responsible for SARS infection. SARS coronavirus 3C-like proteinase (SARS 3CLpro ) plays key roles in viral replication and transcription and is an attractive target for anti-SARS drug discovery. In this report, we quantitatively characterized the dimerization features of the full-length and N-terminal residues 1-7 deleted SARS 3CL pro s by using glutaraldehyde cross-linking SDS-PAGE, size-exclusion chromatography, and isothermal titration calorimeter techniques. Glutaraldehyde cross-linking SDS-PAGE and size-exclusion chromatography results show that, similar to the full-length SARS 3CLpro , the N-terminal deleted SARS 3CL pro still remains a dimer/monomer mixture within a wide range of protein concentrations. Isothermal titration calorimeter determinations indicate that the equilibrium dissociation constant (K d ) of the N-terminal deleted proteinase dimer (262 M) is very similar to that of the full-length proteinase dimer (227 M). Enzymatic activity assay using the fluorescence resonance energy transfer method reveals that N-terminal deletion results in almost complete loss of enzymatic activity for SARS 3CL pro . Molecular dynamics and docking simulations demonstrate the N-terminal deleted proteinase dimer adopts a state different from that of the full-length proteinase dimer, which increases the angle between the two protomers and reduces the binding pocket that is not beneficial to the substrate binding. This conclusion is verified by the surface plasmon resonance biosensor determination, indicating that the model substrate cannot bind to the N-terminal deleted proteinase. These results suggest the N terminus is not indispensable for the proteinase dimerization but may fix the dimer at the active state and is therefore vital to enzymatic activity.

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The complete sequence (22 kilobases) of murine coronavirus gene 1 encoding the putative proteases and RNA polymerase

Cited by 346 publications

References 51 publications

Localization of mouse hepatitis virus open reading frame 1A derived proteins

Localization of mouse hepatitis virus open reading frame 1A derived proteins

Molecular cloning and expression of a spike protein of neurovirulent murine coronavirus JHMV variant c1-2

Severe Acute Respiratory Syndrome Coronavirus 3C-like Proteinase N Terminus Is Indispensable for Proteolytic Activity but Not for Enzyme Dimerization

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